阿托伐他汀对血管紧张素Ⅱ诱导的人肠系膜动脉平滑肌细胞增殖和细胞内NADPH氧化酶-活性氧通路的影响

Effect of atorvastatin on proliferation of human mesenteric artery smooth muscle cells and cellular NADPH oxidase-active oxygen species signal pathway induced by angiotensin Ⅱ

目的观察阿托伐他汀钙对血管紧张素Ⅱ的促人肠系膜动脉平滑肌细胞增殖和其诱导的平滑肌细胞内NADPH氧化酶-活性氧通路的干预作用。方法用组织贴块法培养的第2～4代人肠系膜动脉平滑肌细胞作为实验标本；以四甲基偶氮唑盐（MTT）法比色测定各组平滑肌细胞增殖率；流式细胞仪检测被双氢乙酰乙酸-二氯荧光黄（DCFH—DA）标记的细胞内活性氧；反转录聚合酶链式反应（RT-PCR）方法检测细胞内p22phox蛋白mRNA的表达变化。结果（1）与空白对照组比较，加入血管紧张素Ⅱ药物组的细胞增殖率、细胞内活性氧物质生成量、细胞内p22phoxmRNA的表达均明显增高（P〈0．01）。（2）与浓度为10^-6moL／L血管紧张素Ⅱ孵育组比较，血管紧张素Ⅱ和阿托伐他汀钙共孵育组的细胞增殖率[阿托伐他汀组（0．242±0．018）比血管紧张素Ⅱ组（0．359±0．024），P〈0．01]、细胞内活性氧[阿托伐他汀组（130±29）比血管紧张素Ⅱ组（180±34），P〈0．01]、细胞内p22phox蛋白mRNA的表达[阿托伐他汀组（0．47±0．07）比血管紧张素Ⅱ组（0．71±0．05），P〈0．01]均降低。结论阿托伐他汀钙可以部分抑制血管紧张素Ⅱ的促人肠系膜动脉平滑肌细胞增殖和其诱导的细胞内NADPH氧化酶-活性氧通路的作用。

Objective To observe the effect of atorvastatin on proliferation of human mesenteric artery smooth muscle cells and the activity variations of NADPH oxidase-active oxygen signal pathway induced by angiotensin Ⅱ. Methods Human mesenteric artery smooth muscle cells were isolated and cultured, proliferation rates were measured by MTT assay; reactive oxygen species was detected with flow cytometry; Cellular p22phox mRNA expression was determined by reverse transcription-polymerase chain reaction （RT-PCR）. Results （1） Compared with control group, the cell proliferation rate, reactive oxygen species level and p22phox mRNA expression were significantly increased in angiotensin Ⅱ group （P 〈 0. 01 ）. （2）In atorvastatin plus angiotensin Ⅱ incubation group, the cell proliferation rate（0. 242 ± 0. 018 vs. 0. 359 ± 0. 024, P 〈 0. 01 ）, reactive oxygen species level （ 130 ± 29 vs. 180 ± 34, P 〈 0. 01 ） and cell p22phox mRNA expression （ 0. 47 ± 0. 07 vs. 0. 71 ± 0. 05, P 〈 0. 01 ） were lower than those in angiotensin Ⅱ incubation group. Conclusions Atorvastatin partially inhibits human mesenteric artery smooth muscle cell proliferation and NADPH oxidase- active oxygen signal conduction pathway induced by angiotensin Ⅱ.